Question

Asked 14th Nov, 2015

Ege University

How can I minimize cell stress and decreasing cell adherent during scratch assay on fibroblast cells? How can I optimize scratch assay?

When I triggered scratch assays on Human fibrablast cells, at that time there was no problem; I made wound properly, cells were adherent. I took images for first time point (0h), But, 24 h later there was no adherent cell ( not wound area) in some wells. I always took care about wound, washing ( minimum;once) and aspiration steps.(I tried to minimize all stress factors to effect cells or wound process). But, still, problem is proceeding.

How can I solve this problem?

Thank you for your concern.

Sinem

Scratch Assay

Cell Stress

Cell Culture Techniques

Sinem Yılmaz

Ege University

But, this problem occured not in all well. In few wells cell detaching and stress were seen.

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Anton Vrdoljak

University of Mostar

Malcolm Nobre

Advanced Centre for Treatment, Research and Education in Cancer

Artur Wnorowski

Medical University of Lublin

Antonella Battisti

Italian National Research Council

Ojong Aganyi Asu

University of Calabar

Ru-Jeng Teng

Medical College of Wisconsin

I hope I understand your meaning of "cell swelling". The cells will try to spread out more after scratch so that they can heal off the scratch. So, why is that a concern? Hope I did not make a mistake.

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Sinem Yılmaz

Ege University

Not healing occured. After washing step, cells got stressed and 24 h later numbers of adherent cells got decreased in some wells

Thank you.

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Parthasarathy Chandrakesan

Wild Earth

Fibroblasts need fresh food, change medium often to over come this problem. Cells stressed due to lack of place to divide, lack of food, and decreasing survival due to stress. To avoid such conditions use your scratch analysis when your cells reached 80% confluence and follow with change of medium every 24h, just do the same for your control as for proper comparison.

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2 Recommendations

Sinem Yılmaz

Ege University

But, this problem occured not in all well. In few wells cell detaching and stress were seen.

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1 answer

Asked 2nd Feb, 2018

Sinem Yılmaz

In literature, there are some cell line (dopaminergic neurons, IPS cell lines exc.) in order to make in vitro Parkinson model by neurotoxic agent. One of these cell lines is SHSY-5Y human neuroblastoma cell line. I checked the importance of passage number effect on this Parkinson model; but I found just one research article mentioned that SHSY-5Y cells were used between 7 to 13 passage for Parkinson model. Has anyone have experience about this subject?

Thank you for your concern.

Sincerly.

Sinem

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Thank you for your concern.

Sincerly.

Sinem

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12 answers

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Thanks for your concern.

Sinem

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8 answers

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Question

9 answers

Asked 27th Oct, 2015

Sinem Yılmaz

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Thank you for your concern.

Sinem

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